山西农业大学学报（自然科学版）2019，Vol.39Issue(3)：101-105,5.DOI:10.13842/j.cnki.issn1671-8151.201811048
Cry1Ia和Cry2Ab融合蛋白的表达
Expression of the fusion proteins of cry 1 Iaandcry 2Ab genes
摘要
Abstract
[Objectives]Bacillus thuringiensis (Bt)produces the insecticidal proteins known as Cry toxins such as Cry1Ia and Cry2Ab which hold insecticidal activities against Lepidoptera and Coleoptera pests.In order to take full advantage of thesetox-ins,the fusion proteins of the Cry1Ia and Cry2Ab were constructed and expressed in Bt cells in present study.[Methods]The cry1 Ia gene was fused to the 5'terminus of cry 2Ab gene to generate the cry 1 Ia-cry 2Ab fusion gene.The cry 2Ab-cry 1 Ia fu-sion gene was constructed by similar method except that the c r y 2 Ab genewas located on the 5'terminus.In addition,a trun-cated gene cry 1 IaD 7 3 with 2 1 9 bp deletion in 5'end of the cry 1 Ia gene,which encoding the first α-helix of Cry1 Ia,was ob-tained and inserted into the 5'terminus of cry 2 a bgene.The resulting fusion gene was designated as cry 1 IaD 7 3-cry 2Ab .The three fusion genes were individually inserted into a pHT304 derived plasmid and were controlled by the cry 1 Ac gene promoter and terminator.These vectors were used to express the three fusion proteins in Bt cells.[Results]The SDS-PAGE analysis showed that the three fusion proteins were able to express in Bt cells,and the molecular weight observed were consistent with the calculated sizes,which were 152 kDa,152 kDa,and 144 kDa for cry 1 Ia-cry 2Ab ,cry 2Ab-cry 1 Ia ,and cry 1 IaD 73-c r y 2 Ab ,respectively.Some degradedmolecules of these fusion proteins were detected by western blot analysis.Yields of the target proteins were also affected by the culture temperature.[Conclusion]In this study,three fusion insecticidal genes were constructed and expressed successfully in Bt cells,whichmakes it convenient to investigate theirinsecticidal activities and mode of function in the future.关键词
苏云金芽孢杆菌/cry 1Ia/cry2Ab/融合蛋白/蛋白表达Key words
Bacillus thuringiensis/cry 1 Ia gene/cry 2Ab gene/Fusion protein/Protein expression分类
生物科学引用本文复制引用
郭小琴,钱红梅,郭俊佩,史宗勇,高建华,王兴春..Cry1Ia和Cry2Ab融合蛋白的表达[J].山西农业大学学报（自然科学版）,2019,39(3):101-105,5.基金项目
国家自然科学基金青年基金项目(31201825) （31201825）
山西省科技攻关项目(20150311017-4) （20150311017-4）
